Bovine Dendritic Cell Growth Kit
Bovine dendritic cell growth kit contains a cocktail of biologically active interleukin-4 (IL-4) and granulocyte/macrophage-colony stimulating factor (GM-CSF) that have been premixed at optimal concentrations to induce dendritic cell development from peripheral blood-derived bovine (cattle) monocytes.
- Target Species
- Product Form
- Mixed recombinant bovine Interleukin-4 and bovine GM-CSF – supplied as a liquid
- Recombinant cytokines expressed in mammalian Chinese Hamster Ovary (CHO) cells using the pEE14® vector grown in antibiotic free media and USDA-approved dialysed FCS which has been screened for BVDV and virus growth by PCR.
- Preservative Stabilisers
- None present
- Endotoxin Level
- Store at -20oC only.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature this recombinant protein. Should this product contain a precipitate we recommend microcentrifugation before use.
- Shelf Life
- 6 months from date of despatch.
- This reagent was produced as part of the BBSRC/SEERAD Immunological Toolbox. The kit development was also supported by the European Community’s Seventh Framework Programme (FP7, 2007-2013), Research Infrastructures action, under the grant agreement No. FP7-228394 (NADIR)
- For research purposes only
Applications of Bovine Dendritic Cell Growth Kit
|Application Name||Verified||Min Dilution||Max Dilution|
Where this reagent has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the protein for use in their own system using appropriate negative/positive controls.
- Recommended Protocol
- 1. Prepare peripheral blood mononuclear cells (PBMC) from heparinised blood by density gradient centrifugation.
2. Purify CD14+ve cells by labelling PBMC with CD14 mAb and utilise magnetic bead or flow cytometric separation techniques.
3. Resuspend the isolated CD14+ve cells at a concentration of 1x106 cells/ml in tissue culture medium (TCM = RPMI or equivalent + 10% foetal calf serum) containing a final dilution of 1:20 of PBP015KZZ .
4. Add 3ml of cell suspension to each well of a 6 well tissue culture plate.
5. Culture cells in a humidified atmosphere of 5% CO2 in air, at approximately 37oC.
6. Culture cells for 3 days. The cells may then be harvested and used for other procedures including immunophenotyping (as required).
7. If a longer culture period is required the cells must be ‘fed’ with new TCM containing cytokines on day 3:
Carefully remove 1ml spent medium from each well, care is required to avoid disturbing the cells.
Add 1.5ml fresh, pre-warmed TCM containing cytokines at 1:20 to each well and re-culture the DC for required culture period (typically up to 7 days).
8. At the end of the culture period adherent and non-adherent cells can be pooled for use in immunoassays and phenotyped (as required). Adherent cells may require a dissociation step to remove them from the plate.
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Useful Reagents Available
|Description||Product Code||Pack Size||Applications||List Price||Quantity|
|Mouse anti Human CD14:Alexa Fluor® 647||MCA1568A647||100 Tests/1ml||F|
|Mouse anti Human CD14:Alexa Fluor® 700||MCA1568A700||100 Tests/1ml||F|
|Mouse anti Human CD14:Biotin||MCA1568B||0.1 mg||F|
|Mouse anti Human CD14:Low Endotoxin||MCA1568EL||0.5 mg||E F FN WB|
|Mouse anti Human CD14:FITC||MCA1568F||0.1 mg||F|
|Mouse anti Human CD14:RPE-Alexa Fluor® 647||MCA1568P647||100 Tests/1ml||F|
|Mouse anti Human CD14:Pacific Blue®||MCA1568PB||100 Tests/1ml||F|
|Mouse anti Human CD14:RPE||MCA1568PE||100 Tests||F|
|Mouse anti Bovine CD14:FITC||MCA2678F||0.1 mg||F|
|Mouse anti Bovine MHC Class II DQ||MCA5655||0.25 mg||F IF|
|Mouse anti Bovine MHC Class II DQ:FITC||MCA5655F||0.1 mg||F|
|Mouse anti Bovine MHC Class II DR||MCA5656||0.25 mg||F|
|Mouse anti Bovine CD1w2||MCA831G||0.25 mg||C F IP|
Product Specific References
References for Bovine Dendritic Cell Growth Kit
Hope, J.C. et al. (2000) Dendritic cells induce CD4+ and CD8+ T-cell responses to Mycobacterium bovis and M. avium antigens in Bacille Calmette Guérin vaccinated and nonvaccinated cattle.
Scand J Immunol. 52 (3): 285-91.
Walters, A.A. et al. (2015) Assessment of the enhancement of PLGA nanoparticle uptake by dendritic cells through the addition of natural receptor ligands and monoclonal antibody.
Vaccine. pii: S0264-410X(15)01549-2.
Werling, D. et al. (1999) Involvement of caveolae in the uptake of respiratory syncytial virus antigen by dendritic cells.
J Leukoc Biol. 66 (1): 50-8.